JP2004083419A - Permanent wave treatment method using enzyme and treating agent therefor - Google Patents
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Abstract
Description
【０００１】
【発明の属する技術分野】
本発明は、酵素を用いた毛髪等の新規パーマネントウエーブ処理方法及びその処理剤に関する。
温和な条件にてパーマネントウエーブ処理が可能なために毛髪を傷めることなく、不快臭や手荒れ等を抑えるのに効果的である。
【０００２】
【従来の技術】
毛髪等にパーマネントウエーブ処理をする方法として、従来から一般的に２段階的に実施されている。
第１段階は、システイン、チオグリコール酸又はそれらの塩を用いた還元剤を主成分とする第１剤にて毛髪中のシスチンにおけるジスルフィド結合（Ｓ−Ｓ）を還元開裂させ、ロッド等に巻き付け（ロッド等に巻き付けた後に第１剤を塗布する場合も含む）、その状態にて、新たな位置でジスルフィド結合をさせるべく、臭素酸塩、過酸化水素水等の酸化剤を主成分とする第２剤が用いられている。
【０００３】
上記の場合に第１剤には、還元剤を毛髪内に充分に浸透させるために毛髪が膨潤、軟化するようにｐＨ８．５以上の比較的強いアルカリ性に調整されているものが殆どであった。
従って、このような強いアルカリ性処理剤を用いると、毛髪が傷みやすく、ウエーブ処理時にアンモニア、硫化水素、メルカプタン類等不快臭の発生や手荒れが生じやすかった。
【０００４】
【発明が解決しようとする課題】
本発明は、上記従来技術に有する技術的課題に鑑みて、毛髪の損傷を抑え、皮膚刺激が少ないパーマネントウエーブ処理方法として、タンパク質あるいはペプチドのジスルフィド結合を還元開裂する酵素を用いた、これまでにない新規のパーマネントウエーブ処理方法及びその処理剤の提供を目的とする。
【０００５】
【課題を解決するための手段】
本発明の第１の技術的要旨は、毛髪等の構成タンパク質であるケラチン中に含まれるシスチン結合等のジスルフィド結合（Ｓ−Ｓ）を切断（還元開裂）するのに酵素を用いた酵素学的還元活性を利用する点にある。
このように酵素を用いると、ほぼ、中性に近い領域にて毛髪のパーマネントウエーブ処理が可能であり、従来の強いアルカリ性領域における上記処理に比較して条件が温和であり、毛髪の損傷が少なく、アンモニア、硫化水素、メルカプタン等の不快臭の発生を抑えることができる。
【０００６】
第２の技術的要旨は、上記のような毛髪のタンパク質中のジスルフィド結合を還元開裂するジスルフィドレダクターゼとして、本願出願人が先に発見、発明し、特許出願した新規微生物Ｄ１株（Ｂａｃｔｅｒｉｕｍ Ｄ１）（特願２００１−２６５１８１号）（独立行政法人産業技術総合研究所、特許生物寄託センター、受託番号ＦＥＲＭ Ｐ−１８４８１）を培養することで産生されるジスルフィドレダクターゼＤ１を用いる点にある。
このジスルフィドレダクターゼＤ１は、以下のような酵素学的性質を有している。
（１）基質特異性：タンパク質あるいはペプチドに含まれるＳ−Ｓ結合に対して還元活性を有する。
（２）ケラチンを基質として、ｐＨ６．５〜８．０で反応を行った場合に、最大活性温度は３０℃である。
（３）ケラチンを基質として、３０℃、３０分間反応を行った場合に、至適ｐＨ６．５〜８．０である。
（４）ドデシル硫酸ナトリウム−ポリアクリルアミドゲル電気泳動法による測定で分子量が１５ＫＤａである。
【０００７】
以下、ジスルフィドレダクターゼＤ１についてさらに、詳述する。
図５に、ＳＤＳ−ＰＡＧＥ及び等電点電気泳動により、分子量と等電点を測定した結果を示し、ジスルフィドレダクターゼＤ１の分子量は、約１５Ｄａであり、等電点（ｐＩ）７．２であった。
ジスルフィドレダクターゼＤ１のアミノ基末端側のアミノ酸配列を解析した結果、Ｎ末１５残基が明らかになり、以下にその配列を示す。
−Ｐｒｏ−Ｖａｌ−Ａｓｐ−Ｉｌｅ−Ａｌａ−Ａｌａ−Ａｓｐ−Ｃｙｓ−Ｇｌｕ−Ａｒｇ−Ｌｅｕ−Ｈｉｓ−Ａｌａ−Ｌｅｕ−Ｓｅｒ−ＮＨ２
なお、他のタンパク質との相同性を調査した結果、以下にアミノ酸配列を示すＡｒａｂｉｄｏｐｓｉｓ ｔｈａｌｉａｎａ由来のチオレドキシン様タンパク質と約５０％相同性が認められたものの、他の多くの生物種から得られたチオレドキシンとはほとんど相同性が認められなかった。
（Ａｒａｂｉｄｏｐｓｉｓ ｔｈａｌｉａｎａ由来のチオレドキシン様タンパク質のＮ末アミノ酸配列）
−Ｍｅｔ−Ｖａｌ−Ａｓｐ−Ｉｌｅ−Ｈｉｓ−Ｓｅｒ−Ｔｈｒ−Ｇｌｕ−Ｇｌｕ−Ｐｈｅ−Ｌｅｕ−Ｓｅｒ−Ａｌａ−Ｌｅｕ−Ｓｅｒ−ＮＨ２
図６（Ａ）に、ケラチン基質（ケラチンパウダー）を用いた温度変化に対するジスルフィドレダクターゼＤ１の相対活性を示し、図６（Ｂ）にその温度安定性を示す。
この結果、ジスルフィドレダクターゼＤ１の最大活性温度は、約３０℃であり、４０℃以上に対して不安定であることから、低温特性を有していることが明らかになった。
【０００８】
図７に、ジスルフィドレダクターゼＤ１の至適ｐＨ及びそのｐＨ安定性の調査結果を示す。
図７（Ａ）は、後述するそれぞれの緩衝液を用いて各ｐＨに調整後にケラチン基質の下で、３０分間酵素反応させた相対活性度をグラフに表したものである。図７（Ｂ）は、それぞれの緩衝液中に３０分間ジスルフィドレダクターゼＤ１を放置後に、３０分間酵素反応させて活性度を調査したものである。
活性度はエルマン法を用いて遊離チオールをＤＴＮＢ試薬に呈色反応させ、その吸光度（４１２ｎｍ）を測定した。
その結果、ジスルフィドレダクターゼの至適ｐＨは、６．５〜８．０であり、ｐＨ６．０〜８．５の範囲にてｐＨ安定性を示した。
なお、図７（Ｂ）に示す緩衝液の略号は、次のとおりである。
ＡＡ：ａｃｅｔｉｃ ａｃｉｄ（酢酸）、
ＭＥＳ：２−（Ｎ−ｍｏｒｐｈｏｌｉｎｏ）ｅｔｈａｎｅｓｕｌｆｏｎｉｃ ａｃｉｄ（２−エヌ−モルフォリノ エタンスルフォン酸）、
ＭＯＰＳ：３−（Ｎ−ｍｏｒｐｈｏｌｉｎｏ）ｐｒｏｐａｎｅｓｕｌｆｏｎｉｃ ａｃｉｄ（３−エヌ−モルフォリノ
プロパンスルフォン酸）、
ＴＡＰＳ：Ｎ−［Ｔｒｉｓ（ｈｙｄｒｏｘｙｍｅｔｈｙｌ）−ｍｅｔｈｙｌ］−３−ａｍｉｎｏｐｒｏｐａｎｅｓｕｌｆｏｎｉｃ ａｃｉｄ（エヌ−トリスヒドロキシメチルメチル−３−アミノプロパンスルフォン酸）、
ＣＨＥＳ：２−（Ｎ−ｃｙｃｌｏｈｅｘｙｌａｍｉｎｏ）ｅｔｈａｎｅｓｕｌｆｏｎｉｃ ａｃｉｄ（２−エヌ−シクロヘキシルアミノエタンスルフォン酸）、
ＣＡＰＳ：３−（ｃｙｃｌｏｈｅｘｙｌａｍｉｎｏ）−１−ｐｒｏｐａｎｅｓｕｌｆｏｎｉｃ ａｃｉｄ（３−シクロヘキシルアミノ−１−プロパンスルフォン酸）、
ＧＬＹ：ｇｌｙｃｉｎｅ（グリシン）
【０００９】
ジスルフィドレダクターゼＤ１のＳ−Ｓ結合に対する還元特異性を各種の基質を用いて調査した結果を図８に示す。
この結果、ケラチンの様なタンパク質基質から、グルタチオンの様なペプチド基質まで広範囲に基質特異性を有していることが明らかになった。
【００１０】
【発明の実施の形態】
例えば、図４に示すような、ケラチン含有培養液にて約２０℃、１０〜１４日間、Ｂａｃｔｅｒｉｕｍ Ｄ１（受託番号ＦＥＲＭ Ｐ−１８４８１）を大量培養し、この培養液から遠心分離機により菌体を除去し、その上清液を限外ロ過等により、１０〜２５倍に濃縮したＢａｃｔｅｒｉｕｍ Ｄ１産生酵素ジスルフィドレダクターゼＤ１含有濃縮粗液Ａ（Ｃｒｕｄｅ ｅｎｚｙｍｅ液Ａ）を得た。
また、上記限外ロ過によって得られた約１０倍位程度の濃縮液をさらに陰イオン交換クロマトグラフィー及びゲル濾過クロマトグラフィーにかけて、ケラチン分解酵素プロテアーゼＤ１と、ジスルフィド結合に対して還元活性を示す酵素ジスルフィドレダクターゼＤ１を分離し、ジスルフィドレダクターゼＤ１含有精製液Ｂ（Ｐｕｒｅ ｅｎｚｙｍｅ液Ｂ）を得た。
【００１１】
以上のようにして得られた濃縮粗液Ａ（Ｃｒｕｄｅ ｅｎｚｙｍｅ液Ａ）、精製液Ｂ（Ｐｕｒｅ ｅｎｚｙｍｅ液Ｂ）、及び比較例として、蒸留水をそれぞれ用いて３０℃、３時間、パーマネントウエーブ処理したウエーブ形成状態を図１（写真１）に示す。
パーマネントウエーブ効果は、キルビー法にて測定評価した。
長さ１５〜２０ｃｍの毛髪を準備し、所定の洗浄、乾燥後に２０本を一束とし、キルビーの器具に交互にジグザグにかけ、上記試験液を第１剤として上述したように、３０℃、３時間浸漬放置後に流水で試験液を洗い出し、次に商品名アリスティアウエーブＮ（ＷＮ）ノーマルタイプ（製造元、資生ケミカル株式会社）の第２剤に約３０℃、約１５分間浸漬後に流水洗浄乾燥した。
形成されたウエーブの山５つ分の距離をｂｍｍとし、このｂをまっすぐに伸ばして測定した長さをｃｍｍとする。
キルビーの器具の毛髪をジグザグに交互に巻いた棒の第１番目と第６番目の間の距離をａｍｍとする。
パーマネントウエーブ効果（％）＝１００−１００×（ｂ−ａ）／（ｃ−ａ）
として求められる。
その結果、図１（ハ）に示す蒸留水によるものは全くウエーブを形成しなかったが、図１（イ）濃縮粗液Ａ（Ｃｒｕｄｅ ｅｎｚｙｍｅ液Ａ）、図１（ロ）精製液Ｂ（Ｐｕｒｅ ｅｎｚｙｍｅ液Ｂ）によりウエーブ処理したものは、約７％のパーマネントウエーブ効果を示した。
【００１２】
上記に用いた酵素濃度が精製濃縮後においても、０．０１４ｍｇ／ｍｌ程度の非常に薄いものであったことから、商品名アリスティアウエーブＮ（ＷＮ）ノーマルタイプ（製造元、資生ケミカル株式会社）の第１剤との混合によるウエーブ処理試験を実施した結果を図２（グラフ２）に示す。
横軸がＣｒｕｄｅ ｅｎｚｙｍｅ液Ｂに対する上記市販ウエーブ処理第１剤の混合割合を示し、縦軸が、第１剤で約１５分処理した後に第２剤で約１５分処理したもののパーマネントウエーブ効果（％）を示す。
その結果、Ｃｒｕｄｅ ｅｎｚｙｍｅ液Ｂに対して同量の市販の第１剤を混合した約５０％濃度において、最もジスルフィドレダクターゼＤ１の効果が認められた。
これは、Ｃｒｕｄｅ ｅｎｚｙｍｅ液Ｂだけでは酵素濃度が低く、毛髪をパーマネントウエーブ形成させるのが弱く、また、市販の第１剤の混合割合が高すぎると、酵素による効果が消されてしまうためと推定される。
次に、ジスルフィドレダクターゼＤ１の効果を明確にすべくＣｒｕｄｅ ｅｎｚｙｍｅ液Ｂを一度沸騰させた後に、前述市販第１剤５０％混合したものを比較評価したのが図３（グラフ３）である。
図３（グラフ３）は、第１剤としての処理時間１５分の場合のサンプル３回のパーマネントウエーブ効果（％）の平均値を示す。
これにより、酵素を一度沸騰処理し、酵素効果を消失させたものでも比較例とした、蒸留水に市販の第１剤５０％混合したものより若干パーマネントウエーブ効果が高いものの、沸騰処理しないものより、パーマネントウエーブ効果が低くなっていることから、明らかに酵素によるパーマネントウエーブ効果が確認できた。
【００１３】
【発明の効果】
本発明によれば、ジスルフィド結合に対する還元活性を有する酵素を用いることにより、従来より温和な条件にて毛髪等のパーマネントウエーブ処理が可能となることが明らかになった。
また、これにより、毛髪の損傷を抑え、不快臭の発生を低減し、手荒れ等の取扱い上の問題を抑えたパーマネントウエーブ処理第１剤の提供が期待される。
さらには、タンパク質の改質や、羊毛等のタンパク質系繊維の調質においても本発明に係る酵素の適用が期待される。
【図面の簡単な説明】
【図１】本発明に係る酵素を用いた毛髪のパーマネントウエーブ処理の外観を示す。
【図２】（グラフ２） 濃縮粗液Ａに対する市販第１剤混合効果を示す。
【図３】（グラフ３）酵素の煮沸処理の有無によるパーマネントウエーブ効果比較を示す。
【図４】新規細菌Ｂａｃｔｅｒｉｕｍ Ｄ１の培養液例を示す。
【図５】ジスルフィドレダクターゼＤ１の分子量及び等電点の測定結果を示す。ＫＤａの１が分子量マーカーを示し、２がジスルフィドレダクターゼＤ１の値を示す。
【図６】ジスルフィドレダクターゼＤ１の至適温度調査結果を示す。
【図７】ジスルフィドレダクターゼＤ１の至適ｐＨ調査結果を示す。
【図８】ジスルフィドレダクターゼＤ１の基質特異性を示す。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a novel permanent wave treatment method for hair and the like using an enzyme and a treating agent therefor.
Since the permanent wave treatment can be performed under mild conditions, it is effective in suppressing unpleasant odor and rough hands without damaging the hair.
[0002]
[Prior art]
2. Description of the Related Art As a method of performing a permanent wave treatment on hair or the like, it has been conventionally generally performed in two stages.
In the first step, a disulfide bond (SS) in cystine in hair is reductively cleaved with a first agent mainly containing a reducing agent using cysteine, thioglycolic acid or a salt thereof, and wound around a rod or the like. (Including the case where the first agent is applied after being wound around a rod or the like) In this state, in order to form a disulfide bond at a new position, an oxidizing agent such as bromate or hydrogen peroxide is used as a main component. A second agent is used.
[0003]
In the above case, most of the first agent was adjusted to a relatively strong alkalinity of pH 8.5 or more so that the hair swelled and softened so that the reducing agent could sufficiently penetrate into the hair. .
Therefore, when such a strong alkaline treating agent is used, the hair is easily damaged, and unpleasant odors such as ammonia, hydrogen sulfide, and mercaptans are easily generated and the hands are rough during the wave treatment.
[0004]
[Problems to be solved by the invention]
The present invention has been made in view of the above-mentioned technical problems of the prior art, using an enzyme that reductively cleaves a disulfide bond of a protein or peptide as a permanent wave treatment method that suppresses damage to hair and reduces skin irritation. The purpose of the present invention is to provide a novel permanent wave treatment method and a treating agent therefor.
[0005]
[Means for Solving the Problems]
A first technical gist of the present invention is an enzymatic method using an enzyme to cleave (reductive cleavage) a disulfide bond (SS) such as a cystine bond contained in keratin which is a constituent protein of hair and the like. The point is to use the reducing activity.
By using the enzyme in this way, it is possible to perform permanent wave treatment on the hair in a nearly neutral region, the conditions are milder than those in the conventional treatment in a strongly alkaline region, and the hair is less damaged. And generation of unpleasant odors such as ammonia, hydrogen sulfide and mercaptan.
[0006]
The second technical gist is a novel microorganism D1 strain (Bacterium D1), which was previously discovered, invented, and filed by the present applicant as a disulfide reductase that reductively cleaves a disulfide bond in a hair protein as described above (Bacterium D1) ( (Japanese Patent Application No. 2001-265181) (National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary, Accession No. FERM P-18481) is used to produce disulfide reductase D1.
This disulfide reductase D1 has the following enzymatic properties.
(1) Substrate specificity: has a reducing activity on the SS bond contained in the protein or peptide.
(2) When the reaction is carried out at pH 6.5 to 8.0 using keratin as a substrate, the maximum activity temperature is 30 ° C.
(3) When the reaction is performed at 30 ° C. for 30 minutes using keratin as a substrate, the optimum pH is 6.5 to 8.0.
(4) The molecular weight is 15 KDa as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
[0007]
Hereinafter, the disulfide reductase D1 will be described in more detail.
FIG. 5 shows the results of measuring the molecular weight and isoelectric point by SDS-PAGE and isoelectric focusing. The molecular weight of disulfide reductase D1 was about 15 Da and the isoelectric point (pI) was 7.2. Was.
As a result of analyzing the amino acid sequence on the amino terminal side of disulfide reductase D1, 15 residues at the N-terminal were revealed, and the sequence is shown below.
-Pro-Val-Asp-Ile- Ala-Ala-Asp-Cys-Glu-Arg-Leu-His-Ala-Leu-Ser-NH 2
In addition, as a result of investigating the homology with other proteins, although about 50% homology was recognized with the thioredoxin-like protein derived from Arabidopsis thaliana having the following amino acid sequence, thioredoxin obtained from many other species was obtained. And almost no homology was recognized.
(N-terminal amino acid sequence of thioredoxin-like protein derived from Arabidopsis thaliana)
-Met-Val-Asp-Ile- His-Ser-Thr-Glu-Glu-Phe-Leu-Ser-Ala-Leu-Ser-NH 2
FIG. 6 (A) shows the relative activity of disulfide reductase D1 against a temperature change using a keratin substrate (keratin powder), and FIG. 6 (B) shows the temperature stability.
As a result, the maximum activity temperature of disulfide reductase D1 was about 30 ° C., which was unstable at 40 ° C. or higher.
[0008]
FIG. 7 shows the results of investigation of the optimum pH of disulfide reductase D1 and its pH stability.
FIG. 7 (A) is a graph showing the relative activity obtained by performing an enzymatic reaction under a keratin substrate for 30 minutes after adjusting each pH using each buffer described below. FIG. 7 (B) shows the results obtained by leaving disulfide reductase D1 in each buffer solution for 30 minutes and then performing an enzyme reaction for 30 minutes to investigate the activity.
For the activity, the free thiol was subjected to a color reaction with a DTNB reagent using the Ellman method, and the absorbance (412 nm) was measured.
As a result, the optimum pH of disulfide reductase was 6.5 to 8.0, and showed pH stability in the range of pH 6.0 to 8.5.
The abbreviations of the buffer shown in FIG. 7 (B) are as follows.
AA: acetic acid (acetic acid),
MES: 2- (N-morpholino) ethansulfonic acid (2-N-morpholino ethanesulfonic acid),
MOPS: 3- (N-morpholino) propanesulfonic acid (3-N-morpholinopropanesulfonic acid),
TAPS: N- [Tris (hydroxymethyl) -methyl] -3-aminopropanesulfonic acid (N-trishydroxymethylmethyl-3-aminopropanesulfonic acid),
CHES: 2- (N-cyclohexylamino) ethanesulfonic acid (2-N-cyclohexylaminoethanesulfonic acid),
CAPS: 3- (cyclohexylamino) -1-propanesulfonic acid (3-cyclohexylamino-1-propanesulfonic acid),
GLY: glycine (glycine)
[0009]
FIG. 8 shows the results obtained by examining the reduction specificity of the disulfide reductase D1 for the SS bond using various substrates.
As a result, it was revealed that the substrate has a wide range of substrate specificities from a protein substrate such as keratin to a peptide substrate such as glutathione.
[0010]
BEST MODE FOR CARRYING OUT THE INVENTION
For example, as shown in FIG. 4, a large amount of Bacterium D1 (Accession No. FERM P-18481) is cultured in a keratin-containing culture solution at about 20 ° C. for 10 to 14 days. The resulting supernatant was removed by ultrafiltration or the like to obtain a concentrated crude solution A (Crude enzyme solution A) containing Bacterium D1 producing enzyme disulfide reductase D1 which was concentrated 10 to 25 times.
Further, the concentrated solution of about 10 times obtained by the above-mentioned ultrafiltration is further subjected to anion exchange chromatography and gel filtration chromatography to obtain an enzyme exhibiting a reducing activity for keratinase D1 and a disulfide bond. The disulfide reductase D1 was separated to obtain a purified solution B containing disulfide reductase D1 (Pure enzyme solution B).
[0011]
The concentrated crude liquid A (Crude enzyme liquid A), the purified liquid B (Pure enzyme liquid B) obtained as described above, and, as a comparative example, each were subjected to permanent wave treatment at 30 ° C. for 3 hours using distilled water. FIG. 1 (Photo 1) shows the wave formation state.
The permanent wave effect was measured and evaluated by the Kilby method.
After preparing hair having a length of 15 to 20 cm, after predetermined washing and drying, 20 hairs are bundled, and zigzag alternately is applied to a Kilby apparatus. After immersion for a period of time, the test liquid was washed out with running water, then immersed in a second agent of Aristia Wave N (WN) normal type (manufactured by Shiseido Chemical Co., Ltd.) at about 30 ° C. for about 15 minutes, washed with running water and dried. .
The distance corresponding to five peaks of the formed wave is defined as bmm, and the length measured by extending this b straight is defined as cmm.
Amm is the distance between the first and sixth bars of the zigzag alternately wound hair of the Kilby appliance.
Permanent wave effect (%) = 100-100 × (ba) / (ca)
Is required.
As a result, the sample made of distilled water shown in FIG. 1 (c) did not form any wave, but FIG. 1 (a) concentrated crude solution A (Crude enzyme solution A) and FIG. 1 (b) purified solution B (Pure The one treated with enzyme solution B) showed a permanent wave effect of about 7%.
[0012]
Since the enzyme concentration used above was very thin, about 0.014 mg / ml, even after purification and concentration, the product was manufactured by Aristia Wave N (WN) normal type (manufactured by Shiseido Chemical Co., Ltd.). FIG. 2 (Graph 2) shows the results of a wave treatment test performed by mixing with the first agent.
The horizontal axis indicates the mixing ratio of the commercially available wave treatment first agent to Crude enzyme liquid B, and the vertical axis indicates the permanent wave effect (%) obtained by treating the first agent for about 15 minutes and then treating the second agent for about 15 minutes. ).
As a result, the effect of disulfide reductase D1 was most observed at a concentration of about 50% in which the same amount of the commercially available first agent was mixed with Crude enzyme solution B.
This is presumably because Crude enzyme liquid B alone has a low enzyme concentration, weakly forms a permanent wave on the hair, and if the mixing ratio of the commercially available first agent is too high, the effect of the enzyme is lost. Is done.
Next, in order to clarify the effect of the disulfide reductase D1, the Crude enzyme solution B was boiled once, and then a mixture obtained by mixing 50% of the above-mentioned commercially available first agent was compared and evaluated (FIG. 3).
FIG. 3 (Graph 3) shows the average value of the permanent wave effect (%) of three samples when the treatment time as the first agent is 15 minutes.
As a result, the enzyme was once boiled and the enzyme effect was eliminated. As a comparative example, although the permanent wave effect was slightly higher than that obtained by mixing 50% of the commercially available first agent in distilled water, the enzyme was not boiled. Since the permanent wave effect was low, a permanent wave effect by the enzyme was clearly confirmed.
[0013]
【The invention's effect】
According to the present invention, it has been revealed that permanent waving of hair and the like can be performed under milder conditions by using an enzyme having a reducing activity for disulfide bonds.
In addition, it is expected that the first agent for permanent wave treatment, which suppresses damage to hair, reduces generation of unpleasant odor, and suppresses handling problems such as rough hands, can be provided.
Further, the application of the enzyme according to the present invention is also expected in the modification of proteins and the refining of protein fibers such as wool.
[Brief description of the drawings]
FIG. 1 shows the appearance of a permanent wave treatment of hair using the enzyme according to the present invention.
FIG. 2 (Graph 2) shows the effect of mixing the first agent on the market with concentrated crude liquid A.
FIG. 3 (Graph 3) shows a comparison of a permanent wave effect between the presence and absence of boiling treatment of an enzyme.
FIG. 4 shows an example of a culture solution of the novel bacterium Bacterium D1.
FIG. 5 shows the measurement results of the molecular weight and isoelectric point of disulfide reductase D1. 1 of KDa indicates a molecular weight marker, and 2 indicates the value of disulfide reductase D1.
FIG. 6 shows the results of investigation on the optimum temperature of disulfide reductase D1.
FIG. 7 shows the results of investigation on the optimum pH of disulfide reductase D1.
FIG. 8 shows the substrate specificity of disulfide reductase D1.
Claims (3)
（１）基質特異性：タンパク質あるいはペプチドに含まれるＳ−Ｓ結合に対して還元活性を有する。
（２）ケラチンを基質として、ｐＨ６．５〜８．０で反応を行った場合に、最大活性温度は３０℃である。
（３）ケラチンを基質として、３０℃、３０分間反応を行った場合に、至適ｐＨ６．５〜８．０である。
（４）ドデシル硫酸ナトリウム−ポリアクリルアミドゲル電気泳動法による測定で分子量が１５ＫＤａである。The permanent wave treating agent according to claim 1, wherein the enzyme for cleaving a disulfide bond used in the method for treating permanent wave according to claim 1 is disulfide reductase D1 having the following enzymatic properties.
(1) Substrate specificity: has a reducing activity on the SS bond contained in the protein or peptide.
(2) When the reaction is carried out at pH 6.5 to 8.0 using keratin as a substrate, the maximum activity temperature is 30 ° C.
(3) When the reaction is performed at 30 ° C. for 30 minutes using keratin as a substrate, the optimum pH is 6.5 to 8.0.
(4) The molecular weight is 15 KDa as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
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| JP2002242561A JP2004083419A (en) | 2002-08-22 | 2002-08-22 | Permanent wave treatment method using enzyme and treating agent therefor |
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| JP2002242561A JP2004083419A (en) | 2002-08-22 | 2002-08-22 | Permanent wave treatment method using enzyme and treating agent therefor |
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| JP2004083419A true JP2004083419A (en) | 2004-03-18 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016175577A1 (en) * | 2015-04-30 | 2016-11-03 | 조선대학교 산학협력단 | Perm oxidizing agent composition containing enzyme |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016175577A1 (en) * | 2015-04-30 | 2016-11-03 | 조선대학교 산학협력단 | Perm oxidizing agent composition containing enzyme |
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